Cloning and characterization of a NADP+-malic enzyme gene from Bradyrhizobium japonicum USDA110 |
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| Authors: | Suphawat Sinsuwongwat Amane Kodera Takakazu Kaneko Satoshi Tabata Mika Nomura Shigeysuki Tajima |
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| Affiliation: | 1. Department of Biotechnology, Faculty of Agro-Industry , Chiangmai University , Chiangmai, 50100, Thailand;2. Department of Life Science, Faculty of Agriculture , Kagawa University , Miki-cho, Kita-gun, Kagawa, 761-0795, Japan;3. Kazusa DNA Research Institute , Kisarazu, 292-0812, Japan |
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| Abstract: | Malic enzymes have been considered to play a key role in energy metabolism for nitrogenase reaction in bacteroids. To elucidate the physiological role of the malic enzymes in Bradyrhizobium japonicum bacteroids, a putative malic enzyme gene Bjtme1 was cloned by polymerase chain reaction (PCR) using degenerated primers from conserved regions of the protein sequences of bacterial malic enzymes and draft sequence data of the Bradyrhizobium japonicum USDA110 genome sequence project. To confirm the characteristics of the Bjtme1 gene, the protein encoded by this gene was over-expressed using a pET32a(+) system and it exhibited a NADP+-malic enzyme (EC 1.1.1.40) activity, indicating that Bjtme1 was the gene of the NADP+-malic enzyme. This is the first report on the cloning and characterization of the NADP+-malic enzyme gene from B. japonicum, and the gene structure was compared with that of NADP+-malic enzyme genes of other rhizobia. |
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| Keywords: | Bradyrhizobium japonicum C4-dicarboxylic acid gene cloning NADP+-malic enzymes symbiotic nitrogen fixation |
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